CN107328763A - N NO system and method in a kind of chemical extraction heat energy analytic approach detection Nadroparin Calcium - Google Patents
N NO system and method in a kind of chemical extraction heat energy analytic approach detection Nadroparin Calcium Download PDFInfo
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Abstract
The present invention relates to the system and method for N NO in a kind of chemical extraction heat energy analytic approach detection Nadroparin Calcium, the system includes TEA analyzers, one-level condensing unit, B grade condensing unit, peristaltic pump, heater, reactor, source of oxygen, vavuum pump and source nitrogen.This method includes：S1：Solution is prepared；S2：Instrument regulation etc. prepares；S3：Condensed in two stages device is opened, condensed in two stages is started cooling medium circulation；S4：Partial reaction system, regulation nitrogen flow rate etc. are added from the sample inlet of reactor；S5：Standard solution and need testing solution are added, chromatogram is recorded；Calculate result.The system sets ingenious, simple, can be completely suitable for the detection of NO in Nadroparin Calcium.This method result is more accurate, detection limit is lower, detection speed faster, it is simpler, and this method enormously simplify program, and it has high sensitivity and the high selectivity to nitrogen-containing compound, is the extremely strong detection method of selectivity.
Description
Technical field
The present invention relates to chemical composition quantitative detection field, and in particular to a kind of chemical extraction-heat energy analytic approach detects that
The system and method for bending N-NO in calciparine.
Background technology
The heparin class antithrombotic reagent of new generation that developed recently gets up during LMWHs, its anti thrombotic action is better than liver
Element, and blood coagulation resisting function is less than heparin, and absorb good, bioavilability height, Half-life in vivo length, bleeding with being subcutaneously injected
The advantages of being inclined to small, is widely used to clinic at present.Nadroparin Calcium is one kind of LMWHs, by nitrous acid degradation liver
Element is obtained, average weight-molecular mass 3600~5000, is clinically used for prevention Post operation thromboembolism, prevention of deep vein thrombosis shape
Anti-coagulants, tail vein lesion into extracorporal circulatory system when, pulmonary embolism, haemodialysis etc..In recent years many studies have shown that, nadroparin
The clinical indication of calcium is more and more extensive.
Nitrous acid is used in the production process of Nadroparin Calcium, nitrous acid and heparin generate nitrous under certain conditions
Amines, contains N-NO groups.Substantial amounts of zoopery has confirmed that nitrosamine is strong carcinogen, and can pass through placenta and breast
Juice triggers offspring's tumour.Therefore, the content of N-NO in Nadroparin Calcium is detected, is the content control to nitrosamine compound,
To ensureing that patient medication is significant safely.Cleaning Principle：Sample discharges NO2 and NO, is removed by filtering ＆ cold-traps
Go to crack accessory substance, in the reaction chamber, NO and O3 reacts, and obtains excited electronic state NO2, detects by photomultiplier,
Region of ultra-red emission spectrum, signal is shown finally by integrator.
N-NO groups are mainly detected with thermal energy analyzer in current Nadroparin Calcium.System carrier gas is argon gas, is passed
Bulk cargo puts the middle reagent for needing to use in configuration bubble trap and cold-trap, is filled to after device and is tested, reagent configuration
Process is complicated, and device consuming space is larger, condenser temperature control trouble, and mixing can not be vibrated during heating response system.
The content of the invention
In view of this, it is an object of the invention to provide N- in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium
NO system and method, solve the deficiencies in the prior art.
The purpose of the present invention is to be achieved through the following technical solutions：
N-NO detecting system in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, including TEA analyzers,
Chemiluminescence detector is provided with the TEA analyzers, TEA analyzers are provided with can provide the oxygen of oxygen for TEA analyzers
Air source inlet, the N-NO mixed gas entrance connected with chemiluminescence detector and vacuum pump interface；The detecting system is also wrapped
Include one-level condensing unit, B grade condensing unit, peristaltic pump, heater, reactor, source of oxygen, vavuum pump and source nitrogen.
The one-level condensing unit is provided with cooling medium flow cavity and the first simulation model for mixing gases flows chamber, the cooling medium
Flow cavity is provided with the first condensing medium outlet and the first condensing medium inlet, and the first simulation model for mixing gases flows chamber is provided with the
One mixed gas entrance and the first mixed gas outlet；
Cooling medium storage chamber and the second simulation model for mixing gases flows chamber, the cooling medium are provided with the B grade condensing unit
Storage chamber is provided with the second condensing medium outlet and the second condensing medium inlet, and the second simulation model for mixing gases flows chamber is provided with phase
The second intercommunicated mixed gas entrance and the second mixed gas outlet, the second mixed gas entrance and one-level condensing unit
On the first mixed gas outlet connect so that mixed gas in reactor enters B-grade condensation dress by one-level condensing unit
In putting, second mixed gas outlet is connected by gas circuit pipe with the N-NO mixed gas entrances on TEA；Second condensation
Medium inlet is connected with first condensing medium outlet；
The peristaltic pump, it is with the first condensing medium inlet and the second condensing medium outlet is connected so that B grade condensing unit
In cooling medium enter one-level condensing unit in；
The reactor is provided with gas vent and can add sample and be passed through the sample inlet of nitrogen, and the gas goes out
Mouth is connected with the first mixed gas entrance so that mixed gas in reactor is into one-level condensing unit, the sample inlet
It is connected by nitrogen tube with source nitrogen；
Reactor is placed with the heater；
Oxygen source inlet in the thermal energy analyzer is connected with source of oxygen, and the vacuum pump interface is connected with vavuum pump.
Further, the nitrogen tube is provided with nitrogen partial pressure valve；Gaseous mixture can be adjusted by being provided with the TEA analyzers
The nitrogen adjustment valve of body inlet, the oxygen regulating valve of oxygen inlet.
Further, the one-level condensing unit is condenser pipe；The B grade condensing unit is condenser.
Further, lid of the nitrogen tube on the sample inlet is so as to lead to reactor；The reactor
In be provided with agitating device.
N-NO method in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, according to above-mentioned detecting system
Equipment used in detection is attached, this method comprises the following steps：
S1：Solution is prepared：Need testing solution and standard solution are directly prepared using formamide solution dilution；
S2：1. open vavuum pump to vacuumize TEA analyzers, vacustat opens TEA after below 0.15torr
Analyzer；2. when the vacuum shown by TEA analyzers is kept at below 0.13torr in 20min, nitrogen regulation is opened
Nitrogen partial pressure valve on valve and nitrogen tube, and nitrogen partial pressure is adjusted to 4~6psi；3. oxygen regulating valve is rotated, makes vacuum
0.50 ± 0.01torr is reached, nitrogen partial pressure valve and nitrogen adjustment valve is then rotated, when making partial pressure valve pressure for 5psi, vacuum
Reach 1.67 ± 0.01torr；4. the ozone generator and photometer in TEA analyzers are opened successively；
S3：B grade condensing unit is opened, the temperature of cooling medium is maintained 0-5 DEG C, then opens peristaltic pump, makes one-level cold
Cooling medium circulation in solidifying device and B grade condensing unit；
S4：Ethyl acetate and hydrobromic acid are added from the sample inlet of reactor, makes closed whole system, regulation enters reaction
Nitrogen flow rate in device is 35 ± 0.5ml/min, adjusts heter temperature, is allowed to the state for keeping continually and steadily seething with excitement；
S5：Standard solution and need testing solution are injected separately into reactor, the NO gases of generation are reacted through cold twice
But enter in chemiluminescence detector, record chromatogram；And calculated the data collected, obtain a result.
Further, it is 35ml/min into the nitrogen flow rate in reactor.
Further, in step sl, the specific method of solution preparation is：Need testing solution：Formamide solution is prepared, so
Need testing solution is made in accurately weighed test sample afterwards, plus formamide solution dilution；
Prepare standard items storing solution：(1) standard items storing solution is prepared：Standard items NDPA is taken, absolute ethyl alcohol dissolving, system is added
Into the solution that NDPA concentration is 10 μ g/ml, sealing lucifuge storage；(2) standard items concentrated wiring liquid is prepared：Precision measures standard items deposit
Liquid, is diluted with absolute ethyl alcohol, and NDPA concentrated wiring liquid is made；(3) calibration curve solution：Precision measures standard items concentrated wiring liquid in right amount,
Diluted with formamide solution, it is the gradient concentration solution gradually increased that concentration containing NDPA, which is made,.
Further, the concentration for preparing standard items storing solution is 10 μ g/ml；The concentration of standard items concentrated wiring liquid is
1000ng/ml；Diluting made NDPA gradient concentration solution with formamide solution is respectively：0ng/ml、60ng/ml、
120ng/ml, 180ng/ml solution, wherein, 0ng/ml be formamide solution in be not added with taking standard items concentrated wiring liquid.
Further, the compound concentration of formamide solution is：19ml formamides and 1ml are added in 1g sulfamic acid
Water, shaking makes into homogeneous solution.
Further, in step S4, in reaction vessel magnetic stir bar is added to be stirred, ethyl acetate and hydrobromic acid
Addition is respectively 15ml and 5ml；The temperature adjustment of heter temperature is 60~70 DEG C；Standard solution and need testing solution are every
Secondary sample size is 30~60ul.
The present invention at least has the advantages that：
The invention provides the system and method for N-NO in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium,
The system sets ingenious, simple, clear, can more accurately detect NO, can be completely suitable for the detection of NO in Nadroparin Calcium.
Method of the invention is higher than the detection result stability of conventional method, more accurate, detection limit is lower, detection speed
Faster, it is simpler, and obtained chromatogram peak type is beautiful, collection of illustrative plates baseline is steady so that calculating speed is faster more convenient, error rate
Extremely low, its result of calculation and actual result are basically identical, with extremely important application value；Specifically, the present invention is only needed
50ul sample size, just can be accurately detected the nitrosamine compound of ppb grades of concentration, its detection limit can reach 0.06ppm with
Under.And the reagent configuration process of this method is very simple, enormously simplify program, saves time, cost and manpower；This hair
It is bright that there is high sensitivity and the high selectivity to nitrogen-containing compound, it is the extremely strong detection method of selectivity.
Specifically, 1. use nitrogen stream and nitrogen flow precision control can be significantly improved into accuracy of detection and stability,
And make that testing result is apparent, detection limit is lower.2. it is specific by specific one-level condensing unit and B grade condensing unit etc.
Set and make it that control temperature etc. is more accurate, and significantly improve accuracy and reduce detection limit.3. in solution preparation, this
Invention eliminate committed step in tradition " being dried in vacuo with phosphorus pentoxide ", greatly save the cycle, the time of saving and into
This, and be directly dissolved in formamide solution and can detect the content of wherein NO groups completely.The preparation of standard items storing solution
It is that actual tests have saved time cost without individually preparing in Storage period and preserving type facilitates test of many times.Standard
The preparation of product solution employs calibration curve method, and calibration curve method can more simply and rapidly carry out sample amounts, as a result calculate more
To be accurate, the resultant error scope than conventional method is obviously reduced.In addition, these above-mentioned methods coordinate the reactant in the present invention
System etc. causes the effect of the present invention substantially due to conventional method, and is more suitable for industrial applications.
Brief description of the drawings
Fig. 1 is the structural representation of the detecting system described in the embodiment of the present invention；
Fig. 2 is the chromatogram of embodiment 3 (i.e. experimental group) Plays product described in the embodiment of the present invention；
Fig. 3 is the chromatogram of test sample in embodiment 3 (i.e. experimental group) described in the embodiment of the present invention；
Fig. 4 is the chromatogram of the standard items of contrast groups in embodiment 4 described in the embodiment of the present invention；
Fig. 5 is the chromatogram of the test sample of contrast groups in embodiment 4 described in the embodiment of the present invention.
Fig. 6 is the chromatogram of the change carrier gas described in the embodiment of the present invention；
Fig. 7 is the chromatogram of the increase stream of nitrogen gas described in the embodiment of the present invention；
Fig. 8 is the chromatogram of the reduction stream of nitrogen gas described in the embodiment of the present invention；
Fig. 9 is the chromatogram for being changed into only one of which condensing unit described in the embodiment of the present invention.
1st, heater, 2, reactor, 3, one-level condensing unit, 4, B grade condensing unit, 5, peristaltic pump, 6, TEA analyzers,
7th, source of oxygen, 8, vavuum pump, 9, source nitrogen, 10, nitrogen tube, 11, flexible pipe, 12, gas circuit pipe, 21, gas vent, 22, sample enters
Mouthful, 31, cooling medium flow cavity, the 32, first simulation model for mixing gases flows chamber, 41, cooling medium storage chamber, the 42, second mixed gas
Flow cavity, 61, N-NO mixed gas entrances, 62, vacuum pump interface, 63, oxygen source inlet, 91, nitrogen partial pressure valve, 311, first
Condensing medium outlet, the 312, first condensing medium inlet, the 321, first mixed gas outlet, the 322, first mixed gas entrance,
411st, the second condensing medium inlet, the 412, second condensing medium outlet, the 421, second mixed gas entrance, the 422, second gaseous mixture
Body is exported.
Embodiment
The technical scheme in the embodiment of the present invention is clearly and completely described below, it is clear that described embodiment
It is a part of embodiment of the invention, rather than whole embodiments.The detailed description of embodiments of the invention presented below is simultaneously
The scope of claimed invention is not intended to be limiting, but is merely representative of the selected embodiment of the present invention.Based in the present invention
Embodiment, the every other embodiment that this area commonsense method personnel are obtained under the premise of creative work is not made,
Belong to the scope of protection of the invention.
Embodiment 1
N-NO detecting system in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, as shown in figure 1, including
Chemiluminescence detector is provided with thermal energy analyzer (TEA analyzers) 6, the TEA analyzers 6, TEA analyzers 6 are provided with energy
Oxygen source inlet, the 63 N-NO mixed gas entrances 61 connected with chemiluminescence detector of oxygen are enough provided for TEA analyzers 6
With vacuum pump interface 62, N-NO mixed gas entrance 61 makes N-NO gases in standard items or test sample etc. as nitrogen is from the mouth
Chromatogram record is carried out into TEA；The detecting system also includes one-level condensing unit 3, B grade condensing unit 4, peristaltic pump
5th, heater 1, reactor 2, source of oxygen 7, vavuum pump 8 and source nitrogen 9.
The one-level condensing unit 3 is provided with the simulation model for mixing gases flows chamber 32 of cooling medium flow cavity 31 and first, described cold
Solidifying media flow chamber 31, then can be in the first simulation model for mixing gases flows chamber 32 around the periphery of the first simulation model for mixing gases flows chamber 32 is arranged at
Cooled down；The cooling medium flow cavity 31 is situated between provided with the first condensing medium outlet 311 being interconnected and the first condensation
Matter entrance 312, the first simulation model for mixing gases flows chamber 32 is mixed provided with the first mixed gas entrance 322 and first being interconnected
Close gas vent 321.
The simulation model for mixing gases flows chamber 42 of cooling medium storage chamber 41 and second is provided with the B grade condensing unit 4, it is described cold
Solidifying media storage chamber 41, then can be in the second simulation model for mixing gases flows chamber 42 around the periphery of the second simulation model for mixing gases flows chamber 42 is arranged at
Cooled down；The cooling medium storage chamber 41 is situated between provided with the second condensing medium outlet 412 being interconnected and the second condensation
Matter entrance 411, the second simulation model for mixing gases flows chamber 42 is mixed provided with the second mixed gas entrance 421 and second being interconnected
Gas vent 422 is closed, the second mixed gas entrance 421 passes through the first gaseous mixture on flexible pipe 11 and one-level condensing unit 3
Body outlet 321 connect so that mixed gas in reactor 2 entered by one-level condensing unit 3 in B grade condensing unit 4 the
In two simulation model for mixing gases flows chambers 42, finally enter back into and chromatography etc. is carried out in TEA analyzers 6；Second mixed gas goes out
Mouth 422 is connected by gas circuit pipe 12 with the N-NO mixed gas entrance 61 on TEA analyzers 6, then is mixed with NO's in reactor 2
Nitrogen enters in chemiluminescence detector.Second condensing medium inlet 411 connects with first condensing medium outlet 311
Connect so that the cooling medium returned from one-level condensing unit 3 is circulated back to B grade condensing unit from the second condensing medium inlet 411
In cooling medium storage chamber 41 in 4.
The peristaltic pump 5, it is connected with the first condensing medium inlet 312 and the second condensing medium outlet 412, so that by two
Cooling medium in level condensing unit 4 is pumped into one-level condensing unit 3.
The reactor 2 is provided with gas vent 21 and can add sample and be passed through the sample inlet 22 of nitrogen, described
Gas vent 21 is connected with the first mixed gas entrance, so that the mixed gas in reactor 2 enters the of one-level condensing unit 3
In one simulation model for mixing gases flows chamber；The sample inlet 22 is connected by nitrogen tube with source nitrogen 9, and the nitrogen tube 10 is provided with nitrogen
Qi leel pressure valve 91；The connected mode of source nitrogen 9 and reactor 2 is：Lid of the nitrogen tube 10 on the sample inlet
So that the gas outlet of nitrogen tube 10 is stretched into reactor 2, then nitrogen stream just can enter reactor 2.
Reactor 2 is placed with the heater 1, the heater 1 can heat for reactor 2 and keep certain temperature.
Oxygen source inlet 63 on the TEA analyzers 6 is connected with source of oxygen 7, the vacuum pump interface 62 and vavuum pump 8
Connection.The mixed gas inlet entered from N-NO mixed gas entrance 61 can be adjusted by being additionally provided with the TEA analyzers 6
Nitrogen adjustment valve and the oxygen regulating valve of amount of oxygen that is entered from source of oxygen 7 of regulation and control.
As further preferred embodiment, the one-level condensing unit 3 is preferably condenser pipe；The B-grade condensation dress
It is preferably condenser to put 4.
In the present invention, TEA analyzers are commercially available, and used in the present invention is that Ellutia companies produce, sold
TEA analyzers；Condenser pipe and condenser only need to realize the function of the present invention just can, the condenser pipe and condenser can also
Purchase, such as condenser pipe (producer：Ellutia), condenser (producer：Ellutia).
Embodiment 2
N-NO method in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, this method comprises the following steps：
S1：Equipment used in detection is rationally placed according to the structure of the detecting system described in embodiment 1
And connect；
S2：Solution is prepared：
(1) formamide solution is prepared：Sulfamic acid 1g is weighed, 19ml formamides and 1ml water is added, shaking makes into uniform molten
Liquid.
(2) need testing solution is prepared：Accurately weighed test sample Nadroparin Calcium, plus formamide solution quantitatively dilute and are made often
Containing about Nadroparin Calcium 100mg solution in 1ml.
(3) standard items storing solution is prepared：NDPA (N- nitrosos di-n-propylamine) standard items 100mg is taken, 78.62g is added anhydrous
Ethanol dissolving is configured to primary liquid, then takes primary liquid appropriate, is diluted with absolute ethyl alcohol, and it is the molten of 10 μ g/ml that NDPA concentration, which is made,
Liquid is standard items storing solution, the 5 DEG C of storages of sealing lucifuge.
(4) standard items concentrated wiring liquid：Precision measures standard items storing solution in right amount, is diluted with absolute ethyl alcohol, NDPA concentration is made
For 1000ng/ml solution.
(5) calibration curve solution：Precision measures standard items concentrated wiring liquid in right amount, is diluted with formamide solution, is made and respectively contains
NDPA concentration is 0ng/ml, 60ng/ml, 120ng/ml, 180ng/ml solution, and standard curve concentration unit ng/ml is
Ppb, wherein, 0ng/ml be formamide solution in be not added with taking standard items concentrated wiring liquid.
S2：1. open vavuum pump to vacuumize TEA analyzers 6, vacustat is opened after below 0.13torr
TEA analyzers 6；2. when the vacuum shown by TEA analyzers 6 is kept at below 0.13torr in 20min, nitrogen is opened
Nitrogen partial pressure valve in regulating valve and nitrogen tube, and nitrogen partial pressure is adjusted to 5psi, it is then shut off nitrogen partial pressure valve and nitrogen
Regulating valve；Partial pressure valve pressure, which does not decline, can carry out subsequent step；3. rotate oxygen regulating valve, make vacuum reach 0.50 ±
0.01torr, then rotates nitrogen partial pressure valve and nitrogen adjustment valve, when to make partial pressure valve pressure be 5psi, and vacuum reaches 1.67 ±
0.01torr；4. the ozone generator and photometer in TEA analyzers are opened successively；
S3：B grade condensing unit is opened, the temperature of cooling medium is maintained 0-5 DEG C, then opens peristaltic pump, makes one-level cold
Cooling medium circulation in solidifying device and B grade condensing unit；
S4：The lid of sample inlet is outwarded winding, magnetic stir bar is added into reaction vessel, is added from the sample inlet of reactor
Enter 15ml ethyl acetate and 5ml hydrobromic acids, screw sample inlet lid, then closed whole system is determined with gas flowmeter
Into the nitrogen flow rate in reactor, and make nitrogen 35 ± 0.5ml/min of tasselled, the control of nitrogen flow can be directly influenced
Precision is tested, this method preferably enters the nitrogen flow rate 35ml/min in reactor, and regulation heter temperature is 60~70 DEG C,
And stirring rotor speed is adjusted to moderate, to make the holding of the solution in reactor continually and steadily seethe with excitement state；
S5：The standard solution of each concentration is injected separately into reactor, the standard items of each concentration gradient it is parallel enter
The pin of sample 3, the sample size 50ul per pin, after standard solution sample introduction is finished, need testing solution is also injected into reactor every time,
The parallel sample introduction of need testing solution 2 times, each 50ul.
Standard solution and need testing solution with the solution reaction in reactor and generating NO gases, the NO gases with
Nitrogen flow through one-level condensing unit and B grade condensing unit this cool down twice after enter and detected in chemiluminescence detector, note
Record chromatogram；And calculated the data collected, obtain a result.
Embodiment 3
N-NO method in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, this method comprises the following steps：
S1：Equipment used in detection is rationally placed according to the structure of the detecting system described in embodiment 1
And connect；
S2：Solution is prepared：
(1) formamide solution is prepared：Sulfamic acid 1g is weighed, 19ml formamides and 1ml water is added, shaking makes into uniform molten
Liquid.
(2) need testing solution is prepared：Accurately weighed test sample Nadroparin Calcium, plus formamide solution quantitatively dilute and are made often
Containing about Nadroparin Calcium 100mg solution in 1ml.The actual content of N-NO groups is in the need testing solution pre-established
0.065ppm；
(3) standard items storing solution is prepared：NDPA (N- nitrosos di-n-propylamine) standard items 100mg is taken, 78.62g is added anhydrous
Ethanol dissolving is configured to primary liquid, then takes primary liquid appropriate, is diluted with absolute ethyl alcohol, and it is the molten of 10 μ g/ml that NDPA concentration, which is made,
Liquid is standard items storing solution, the 5 DEG C of storages of sealing lucifuge.
(4) standard items concentrated wiring liquid：Precision measures standard items storing solution in right amount, is diluted with absolute ethyl alcohol, NDPA concentration is made
For 1000ng/ml solution.
(5) calibration curve solution：Precision measures standard items concentrated wiring liquid in right amount, is diluted with formamide solution, is made and respectively contains
NDPA concentration is 0ng/ml, 60ng/ml, 120ng/ml, 180ng/ml solution, and standard curve concentration unit ng/ml is
Ppb, wherein, 0ng/ml be formamide solution in be not added with taking standard items concentrated wiring liquid.
S2：1. open vavuum pump to vacuumize TEA analyzers 6, vacustat is opened after below 0.13torr
TEA analyzers 6；2. when the vacuum shown by TEA analyzers 6 is kept at below 0.13torr in 20min, nitrogen is opened
Nitrogen partial pressure valve in regulating valve and nitrogen tube, and nitrogen partial pressure is adjusted to 5psi, it is then shut off nitrogen partial pressure valve and nitrogen
Regulating valve；Partial pressure valve pressure, which does not decline, can carry out subsequent step；3. oxygen regulating valve is rotated, vacuum is reached
0.50torr, then rotates nitrogen partial pressure valve and nitrogen adjustment valve, and when making partial pressure valve pressure for 5psi, vacuum reaches 1.67；
4. the ozone generator and photometer in TEA analyzers are opened successively；
S3：B grade condensing unit is opened, the temperature of cooling medium is maintained 2-3 DEG C, then opens peristaltic pump, makes one-level cold
Cooling medium circulation in solidifying device and B grade condensing unit；
S4：The lid of sample inlet is outwarded winding, magnetic stir bar is added into reaction vessel, is added from the sample inlet of reactor
Enter 15ml ethyl acetate and 5ml hydrobromic acids, screw sample inlet lid, then closed whole system is determined with gas flowmeter
Into the nitrogen flow rate in reactor, and make nitrogen tasselled 35ml/min, regulation heter temperature is 65 DEG C, and stirrer is turned
Velocity modulation makes the solution in reactor keep the state continually and steadily seethed with excitement to moderate；
S5：The standard solution of each concentration is injected separately into reactor, the standard items of each concentration gradient it is parallel enter
The pin of sample 3, the sample size 50ul per pin, after standard solution sample introduction is finished, need testing solution is also injected into reactor every time,
The parallel sample introduction of need testing solution 2 times, each 50ul.
S6：As a result：As shown in Figure 2, sample chromatogram figure is as shown in Figure 3 for resulting standard items chromatogram.
As shown in Figure 2, the peak area and average value of standard items are as shown in table 1 below.Due to standard concentration be respectively 0ng,
60ng, 120ng, 180ng, each concentration distinguish the parallel pin of sample introduction three, there are three peak areas, and average value is to take three times averagely
Value is calculated.
The peak area of the standard items of table 1
N-NO cubage formula are：Standard items peak area and concentration calculate equation of linear regression:CR=kAR+b；Will
Sample peak area substitutes into above equation of linear regression and calculates NDPA concentration Cs in sample solutionS(ng/ml)；Further according to public affairs
Formula CS* 44*V/m/130 show that N-NO group contents (ppm) in test sample, wherein m are sample sample weighting amount, mg；V is sample solution
Volume, ml.
Equation of linear regression can be drawn by the result of table 1, be：C=0.0016A-92.8196；R=0.99.
Sample chromatogram peak as shown in Figure 3 understands that its peak area is:Peak 1：61956；Peak 2：79814, average value is
70885, calculated by formula, it is 0.07ppm to obtain N-NO group contents in test sample (i.e. sample).
Embodiment 4
Experimental group：Method and resulting result described in embodiment 3；The N-NO in embodiment 3, test sample
The actual content of group is 0.065ppm, and it is 0.07ppm that experimental group (i.e. embodiment 3) method, which calculates obtained content, with reality
Border content difference is minimum, as a result substantially close to actual result.
Contrast groups：Standard solution and need testing solution are traditionally prepared, standard concentration is single concentration, is prepared
Concentration etc. is identical with experimental group, and the content of N-NO groups is similarly 0.065ppm in need testing solution；Method of testing is also using biography
System method.The chromatogram of the standard solution of gained is as shown in figure 4, the chromatogram of sample is as shown in Figure 5.Calculate gained standard items
Peak area be：28256, the peak area of sample is 10040, and N-NO in obtained sample is calculated using the formula in embodiment 3
Group content is 0.1ppm.Because actual content is 0.065ppm, and the content that the contrast groups are obtained is 0.1ppm, difference
0.35ppm, the unstability of this method etc. in itself causes it to be differed greatly with actual content, as a result with actual result uniformity very
Difference.
From the chromatogram obtained by Fig. 2~3, the inventive method, either standard items or sample, its peak shape mark
Standard, multiple small peaks of each peak without fission, collection of illustrative plates baseline is steady, and peak area is easily determined, calculates and is very easy to and the degree of accuracy
Height, staff, which only needs to a few minutes, just can easily draw final result, and the result that different staff is calculated is basic
Unanimously, error rate is minimum.
From Fig. 4~5, the collection of illustrative plates baseline that conventional method experiment is drawn is unstable, and peak type is ugly, is split at the top of each peak
Become and multiple small peaks, and be difficult to judge the starting and ending section at the peak, be the difficulty that staff causes calculating, be difficult to sometimes
The quantity at peak is differentiated, and sometimes a sample needs to calculate most of the day, and the degree of accuracy is not high, error rate is higher, different works
Make the result difference that personnel are calculated larger, there is larger error for the presentation of final result.
It is exactly baseline noise than larger that above-mentioned baseline is unstable, baseline noise can interference detection results, if sample is dense
If degree is low, it will not come out with noise point, and calculating of the baseline to peak area has great interference effect.Therefore, put down
Steady baseline is very big for accurate result and low test limit influence.
Therefore, final result of the invention is far superior to conventional method.
In addition it is also necessary to explanation, for the method in embodiment 3, if carrier gas is changed into argon gas, nitrogen is increasedd or decreased
The air-flow of gas, condensed in two stages is changed into only any of which condensing unit etc., larger, chromatogram is influenceed on final result
The effect of figure also has significant change, as shown in accompanying drawing 6~9, and Fig. 6 uses argon gas for carrier, and Fig. 7 and Fig. 8 are respectively increase air-flow
With reduction air-flow, Fig. 9 is is changed to comprise only a condensing unit, from these figures, and these factors compare for spectrogram influence
It is larger, change that any one factor may can all make that spectrogram baseline is unstable, peak fission etc., be the later stage calculating and
As a result accuracy causes many influences.And the present invention collects these improvement together in the present invention, these are improved at that
Under the influence of this so that result is very pleasantly surprised, spectrogram is extremely beautiful, calculates very convenient, error rate is extremely low.The present inventor is from originally
The place that industry research person does not notice that or considered substantially is started with, and by multiple local improvement, has finally given splendid effect
Really, baseline is steady, and calculated by peak area is convenient, accurate, result precision is high.
The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, for those skilled in the art
For, the present invention can have various changes and change.It is all any modifications made within spirit and principles of the present invention, equivalent
Replace, improve etc., it should be included in the scope of the protection.
Claims (10)
1. N-NO detecting system in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, including TEA analyzers, institute
State and chemiluminescence detector is provided with TEA analyzers, TEA analyzers are provided with can provide the oxygen of oxygen for TEA analyzers
Source inlet, the N-NO mixed gas entrance connected with chemiluminescence detector and vacuum pump interface；It is characterized in that：The detection
System also includes one-level condensing unit, B grade condensing unit, peristaltic pump, heater, reactor, source of oxygen, vavuum pump and nitrogen
Source；
The one-level condensing unit is provided with cooling medium flow cavity and the first simulation model for mixing gases flows chamber, the cooling medium flowing
Chamber is provided with the first condensing medium outlet and the first condensing medium inlet, and the first simulation model for mixing gases flows chamber is mixed provided with first
Close gas access and the first mixed gas outlet；
Cooling medium storage chamber and the second simulation model for mixing gases flows chamber, the cooling medium storage are provided with the B grade condensing unit
Chamber is provided with the second condensing medium outlet and the second condensing medium inlet, and the second simulation model for mixing gases flows chamber is provided with and mutually interconnected
On the second logical mixed gas entrance and the second mixed gas outlet, the second mixed gas entrance and one-level condensing unit
First mixed gas outlet is connected so that mixed gas in reactor is entered in B grade condensing unit by one-level condensing unit,
Second mixed gas outlet is connected by gas circuit pipe with the N-NO mixed gas entrances on TEA analyzers；Described second is cold
Solidifying medium inlet is connected with first condensing medium outlet；
The peristaltic pump, it is connected so that in B grade condensing unit with the first condensing medium inlet and the second condensing medium outlet
Cooling medium enters in one-level condensing unit；
The reactor provided with gas vent and sample being added and the sample inlet of nitrogen is passed through, the gas vent with
First mixed gas entrance is connected so that the mixed gas in reactor enters in one-level condensing unit, and the sample inlet passes through
Nitrogen tube is connected with source nitrogen；
Reactor is placed with the heater；
Oxygen source inlet in the thermal energy analyzer is connected with source of oxygen, and the vacuum pump interface is connected with vavuum pump.
2. N-NO detecting system in chemical extraction according to claim 1-heat energy analytic approach detection Nadroparin Calcium, its
It is characterised by：The nitrogen tube is provided with nitrogen partial pressure valve；Mixed gas inlet can be adjusted by being provided with the TEA analyzers
Nitrogen adjustment valve, the oxygen regulating valve of oxygen inlet.
3. N-NO detecting system in chemical extraction according to claim 2-heat energy analytic approach detection Nadroparin Calcium, its
It is characterised by：The one-level condensing unit is condenser pipe；The B grade condensing unit is condenser.
4. N-NO detecting system in chemical extraction according to claim 3-heat energy analytic approach detection Nadroparin Calcium, its
It is characterised by：Lid of the nitrogen tube on the sample inlet is so as to lead to reactor；It is provided with the reactor
Agitating device.
5. N-NO method in a kind of chemical extraction-heat energy analytic approach detection Nadroparin Calcium, it is characterised in that：Will according to right
The detecting system any one of 1~4 is asked to be attached equipment used in detection, this method comprises the following steps：
S1：Solution is prepared：Need testing solution and standard solution are directly prepared using formamide solution dilution；
S2：1. open vavuum pump to vacuumize TEA analyzers, vacustat opens TEA analyses after below 0.15torr
Instrument；2. when the vacuum shown by TEA analyzers is kept at below 0.13torr in 20min, open nitrogen adjustment valve and
Nitrogen partial pressure valve in nitrogen tube, and nitrogen partial pressure is adjusted to 4~6psi；3. oxygen regulating valve is rotated, vacuum is reached
0.50 ± 0.01torr, then rotates nitrogen partial pressure valve and nitrogen adjustment valve, and when making partial pressure valve pressure for 5psi, vacuum reaches
1.67±0.01torr；4. the ozone generator and photometer in TEA analyzers are opened successively；
S3：B grade condensing unit is opened, the temperature of cooling medium is maintained 0-5 DEG C, then opens peristaltic pump, makes one-level condensation dress
Put and the cooling medium circulation in B grade condensing unit；
S4：Ethyl acetate and hydrobromic acid are added from the sample inlet of reactor, makes closed whole system, regulation enters in reactor
Nitrogen flow rate be 35 ± 0.5ml/min, adjust heter temperature, be allowed to the state for keeping continually and steadily seething with excitement；
S5：Standard solution and need testing solution are injected separately into reactor, react generation NO gases through cool down twice into
Enter in chemiluminescence detector, record chromatogram；And calculated the data collected, obtain a result.
6. N-NO method, its feature in chemical extraction according to claim 5-heat energy analytic approach detection Nadroparin Calcium
It is：It is 35ml/min into the nitrogen flow rate in reactor.
7. N-NO method, its feature in chemical extraction according to claim 6-heat energy analytic approach detection Nadroparin Calcium
It is：In step sl, the specific method of solution preparation is：
Need testing solution：Preparing formamide solution, then accurately weighed test sample, plus formamide solution dilution, that test sample is made is molten
Liquid；
Prepare standard items storing solution：
(1) standard items storing solution is prepared：Standard items NDPA is taken, absolute ethyl alcohol dissolving is added, it is 10 μ g/ml's that NDPA concentration, which is made,
Solution, sealing lucifuge storage；
(2) standard items concentrated wiring liquid is prepared：Precision measures standard items storing solution, is diluted with absolute ethyl alcohol, and NDPA concentrated wiring liquid is made；
(3) calibration curve solution：Precision measures standard items concentrated wiring liquid in right amount, is diluted with formamide solution, concentration containing NDPA is made
For the gradient concentration solution gradually increased.
8. N-NO method, its feature in chemical extraction according to claim 7-heat energy analytic approach detection Nadroparin Calcium
It is：The concentration for preparing standard items storing solution is 10 μ g/ml；The concentration of standard items concentrated wiring liquid is 1000ng/ml；Use formyl
Amine aqueous solution dilutes made NDPA gradient concentration solution：0ng/ml, 60ng/ml, 120ng/ml, 180ng/ml's is molten
Liquid, wherein, 0ng/ml be formamide solution in be not added with taking standard items concentrated wiring liquid.
9. N-NO method, its feature in chemical extraction according to claim 8-heat energy analytic approach detection Nadroparin Calcium
It is：The compound concentration of formamide solution is：19ml formamides and 1ml water are added in 1g sulfamic acid, shaking makes into equal
Even solution.
10. N-NO method in chemical extraction according to claim 9-heat energy analytic approach detection Nadroparin Calcium, it is special
Levy and be：Magnetic stir bar is added in step S4, in reaction vessel to be stirred, the addition difference of ethyl acetate and hydrobromic acid
For 15ml and 5ml；The temperature adjustment of heter temperature is 60~70 DEG C；Standard solution and each sample size of need testing solution
It is 30~60ul.
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